Ultrafast photoreduction dynamics of a new class of CPD photolyases

NewPHL is a recently discovered subgroup of ancestral DNA photolyases. Its domain architecture displays pronounced differences from that of canonical photolyases, in particular at the level of the characteristic electron transfer chain, which is limited to merely two tryptophans, instead of the classical three or four. Using transient absorption spectroscopy, we show that the dynamics of photoreduction of the oxidized FAD cofactor in the NewPHL begins similarly as that in canonical photolyases, i.e., with a sub-ps primary reduction of the excited FAD cofactor by an adjacent tryptophan, followed by migration of the electron hole towards the second tryptophan in the tens of ps regime. However, the resulting tryptophanyl radical then undergoes an unprecedentedly fast deprotonation in less than 100 ps in the NewPHL. In spite of the stabilization effect of this deprotonation, almost complete charge recombination follows in two phases of similar to 950 ps and similar to 50 ns. Such a rapid recombination of the radical pair implies that the first FAD photoreduction step, i.e., conversion of the fully oxidized to the semi-quinone state, should be rather difficult in vivo. We hence suggest that the flavin chromophore likely switches only between its semi-reduced and fully reduced form in NewPHL under physiological conditions. [GRAPHICS] .

Automated summary

NewPHL, a newly discovered subgroup of ancestral DNA photolyases, exhibits differences in domain architecture and photoreduction dynamics compared to canonical photolyases. The rapid deprotonation of the resulting tryptophanyl radical in NewPHL leads to quick charge recombination, making the first FAD photoreduction step challenging in vivo. It is suggested that under physiological conditions, the flavin chromophore likely toggles between its semi-reduced and fully reduced form in NewPHL.